Natural preservatives and antimicrobial agents, including compositions thereof

ABSTRACT

Compositions are disclosed herein that comprise a mixture of at least one  Albizia  extract and a  Magnolia  extract. In some embodiments, the composition comprises a mixture of at least one  Albizia  extract enriched for one or more macrocyclic alkaloids and at least one  Magnolia  extract enrich for one or more lignans.

This United States Utility Application claims priority to U.S.Provisional Patent Application Ser. No. 62/252,987 filed on Nov. 9, 2015and entitled “Natural Preservatives and Antimicrobial Agents”, which iscommonly-owned and incorporated herein by reference in its entirety.

FIELD OF THE SUBJECT MATTER

The field of the subject matter relates to compositions that can act aspreservatives, antimicrobial agents, or a combination thereof andrelated methods. Contemplated embodiments are directed to preservativesderived from extracts that are derived from Albizia amara seeds andMagnolia officinalis barks, which can be used to preserve food,cosmetics, nutraceuticals, oral hygiene products, and pharmaceuticals.

BACKGROUND

A preservative is a substance that is added to products, such as foods,pharmaceuticals, cosmetics, biological samples, etc. to preventdecomposition by microbial growth or by undesirable chemical changes.Anti-microbial preservatives prevent the degradation by bacteria, andinhibit the growth of bacteria, fungi and other microorganisms to keepthe product quality and extend the shelf life. A preservative can beused by itself or it can be combined with other physical techniques forpreservation including dehydration, UV-C radiation, freeze-drying,refrigeration, etc. Anti-oxidant preservatives are commonly used in thefood industry, especially the food with a high fat content. Commonanti-oxidants include phenol derivatives BHT (butylated hydroxy toluene)and BHA (butylated hydroxyl anisole). Other preservatives includeformaldehyde, glutaraldehyde, ethanol and methylchloroisothiazolinone.

Chemical preservatives and physical preservation are usually combined.Common anti-microbial preservatives include sodium nitrate, sodiumnitrite, sulfites (sulfur dioxide, sodium bisulfite, and potassiumhydrogen sulfite), benzoic acid/sodium benzoate, sorbic acid/sodiumsorbate, propionic acid/calcium propionate/sodium propionate, anddisodium EDTA targeting on different products. The efficacy, safety andtoxicity of many synthetic preservatives are under the scrutiny amongacademics and regulation agencies because of the adverse reports andconcerns from customers, for instance paraben and its analogues, whichhave been banned in Europe and some Asian countries as well. Naturalpreservatives are considered safer and more environmentally-friendlyalternatives for microbial infection with a broad range of structurallydiversified bioactive compounds such as alkaloids, flavonoids, aromaticterpenoids and saponins.

Natural substances such as salt, sugar, vinegar, tea extracts, herbessential oils and organic acids are well accepted as traditionalpreservatives. For example, citric acid and ascorbic acids from lemon orother citrus juices are commonly used as anti-browning and anti-oxidantin the fruit and juice industry to improve the food quality. However,the current natural preservatives can be expensive or have less potentefficacy, which limits their broader use in food, cosmetics and otherbiological products industry, compared to the synthetic preservatives,especially the antimicrobial preservatives. Efficient, targeted andappropriate preservatives are critical to extend product shelf life,keep product quality, benefit in many aspects in the whole process andtransportation to improve the product safety. Therefore, the idea ofdiscovering and utilizing a special natural composition with potentantimicrobial activity against broad spectrum of microorganisms ispractical and appealing.

Thus, there is a need for alternative natural preservatives andantimicrobial agents. To develop natural preservatives, ahigh-throughput anti-E. coli assay was carried out to screen a plantlibrary of over 10,216 organic extracts (OE) and aqueous extracts (AE),which are generated from medicinal plants collected from all over theworld, Albizia amara seeds extract was identified as the best leadexhibiting potent anti-Gram negative inhibition with a MIC value of 40ug/mL. Bioassay-guided isolation and identification confirmed the activecomponents are budmunchimaine C, as shown in FIG. 1, and its analogues.

Albizia is a genus of about over 150 species distributed mostly insubtropical and tropical areas in Asia, Africa, Madagascar, America andAustralia, but mostly in the old world tropics. Many species has beenused as folk medicines in India and South Africa countries for cold,diarrhea, headache, intestinal ailments, stomach cancer, sore throat andstomachache. For example the stem bark of A. schimperiana is usedindigenously for treating bacterial and parasitic infections, and morecommonly against fever and in pain relief.

Modern studies of these plants showed antimicrobial, antiparasitic,antitrypanosomal and mosquito larvicidal activities. Furtherinvestigation of these Albizia plants leads to the discovery of a uniqueclass of macrocyclic alkaloids, represented by budmunchiamines A-I andother analogues (Dixit 1995; Assis 1999; Misra, 1995; Ovenden 2002;Samoylenko 2009). This characteristic type of macrocyclic alkaloids havebeen reported from seeds of Albizzia amara, Albizzia lebbek, stem barksor barks of Albizzia gummifera, Albizzia schimperana, Albiziasaman/Samanea saman, Albizia schimperiana, Albizia adinocephala leavesof Albizzia inopinata, Samanea saman (Pithecolobium saman=Albizziasaman) with a variety of anti-microbial, anti-parasitic and anticanceractivities (Samoylenko 2009; Geyid 2005; Thippeswamy 2015; March 1991;Pezzutoa 1992).

Magnolia officinalis, commonly known as “houpu” in Chinese, is one ofthe popular components of traditional Chinese medicine having a verywide range of applications is a species of Magnolia native in China,mainly growing in Sichuan and Hubei province. Houpu refers to its thickbark, which could be stripped from the stems, branches, and roots. Thetraditional indications are to treat wind stroke, cold damage,headaches, fight qi and blood impediment. Magnolia bark has been used totreat menstrual cramps, abdominal pain, abdominal bloating and gas,nausea, and indigestion. The bark is also an ingredient in formulas usedfor treating coughs and asthma. Many of the formulations with Magnoliabark are used in treating lung diseases such as including cough andasthma or intestinal infections and spasms, relieving abdominal swellingof various causes and edema.

Modern researches discovered potent antimicrobial and antifungalactivity of this herb medicine against a variety of microorganism, andbisphenolic lignans are identified as the major active componentsresponsible the antimicrobial efficacy. Magnolol and honokiol, as shownin FIG. 2, are two main polyphenol compounds found in the magnolia barksand have been reported with various pharmacological activities andfunctions such as antioxidant, anti-inflammatory, and antitumor (Park2004). The anticancer studies of honokiol have been extended to severaldifferent solid tumor types such as breast, prostate, gastric, andovarian cancer, with potential to enhance current anticancer regimens(Fried 2009). Honokiol also reduce inflammation and oxidative stress,providing beneficial effects in neurological protection, and glucoseregulation with great potential as therapeutic agents for inflammatorydisease. In particular, magnolol and honokiol have been known exhibitingpotent antimicrobial activity against Gram-positive and Gram-negativebacteria as well as fungi such as Propionibacterium sp. and S. aureusshowing its potential as antimicrobial agents effective against moreinfectious resistant microorganisms (Ho 2001; Bang 2000; Kim 2015). Thecontent of honokiol and magnolol could be varied from 1-99% in thecommercialized magnolia bark extracts.

There are a number of references that help provide context as to whysome of these components may be considered important. U.S. Pat. No.8,329,095 discloses a preservative composition for cosmetic formulationscomprising thymol, monolaurin and magnolol obtained from supercriticalfluid extracts of Magnolia officinalis. U.S. Pat. No. 7,592,025discloses a vehicle and a method for oral care that includes MagnoliaBark Extract in combination with a surface-active agent.

CN 101516364 discloses compositions comprising hydroxytyrosol alcoholand/or oleuropein and at least one additional component selected fromthe group of the additional components such ligustilide oleuropeinaglycone, Magnolol, and Park phenol, genistein, resveratrol, EGCG,Magnolia Bark Extract, cashew fruit extract and Glycyrrhiza foetidagroup, contemplated embodiments also relate to the use as apharmaceutical composition, especially as for the treatment of disordersthe treatment or prevention of inflammatory medicine.

CN 1103756 discloses a natural antiseptic agent is made up by mixing90-99.7% of the extractive powder of bark of official magnolia and therest, the extractive powder of Chinese rhubarb.

CN 100544597 discloses a Chinese herbal preservatives of Magnolia,Clematis, Humulus, rugosa as raw materials, based on the differentcharacteristics of each flavor of Chinese herbal medicine.

CN 103416480 discloses a method for a plant source of complex Orangepreservatives, plant extracts, food preservatives, stabilizers andacidity regulating agents composite material. The main technicalfeatures: the main ingredient containing preservative Magnolia extract10-15%, 20-30% clove extract, potassium sorbate 10-20%, 10-20% propylparaben, 2-5% citric acid, 1-5% stabilizer traits as a tan emulsion.

WO 2006069209 discloses an efficacious antibacterial andanti-inflammatory oral composition is provided having an activeingredient combination comprising one or more active compounds from anextract of magnolia and an extract of hops.

WO 2014131191 discloses a composition comprising an antimicrobial activecomprising honokiol and/or magnolol and a carboxylic acid. Also providedare methods of use thereof. None of these references, however, addressthe need in the art for a composition that acts as a preservative,antimicrobial agent or a combination thereof.

SUMMARY OF THE DISCLOSURE

Compositions are disclosed herein that comprise a mixture of at leastone Albizia extract and a Magnolia extract. In some embodiments, thecomposition comprises a mixture of at least one Albizia extract enrichedfor one or more macrocyclic alkaloids and at least one Magnolia extractenrich for one or more lignans.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1. Chemical structure of budmunchiamine C

FIG. 2. Chemical structures of magnolol and honokiol

FIG. 3. Colony counts for 416-145-50 at 1 mg/mL

FIG. 4. Colony counts for 416-145-50 at 500 ug/mL

FIG. 5 Colony counts for 1940601 in ionic o/w emulsion system at 0.5%

FIG. 6 Colony counts for 1940602 in ionic o/w emulsion system at 0.1%

DETAILED DESCRIPTION

In brief, this disclosure and contemplated embodiments are directed tocompounds and compositions which can be used as preservatives andanti-microbial agents, including stereoisomers, pharmaceutically ornutraceutically acceptable salts, tautomers, glycosides and prodrugs ofthe disclosed compounds, and to related methods. Contemplatedembodiments relate to preservatives derived from extracts derived fromAlbizia amara seeds and Magnolia officinalis barks, which can be used topreserve food, cosmetics, nutraceuticals, oral hygiene products, andpharmaceuticals.

Contemplated compounds and compositions are derived from or comprise atleast one plant extract, wherein the plant extract may or may not beenriched. As part of this development, frequently and acceptable assayswere utilized to test contemplated compounds and compositions.

Specifically, compositions, compounds and methods for preservatives andantimicrobial compositions are disclosed that include a mixture of plantextracts, wherein the plant extracts comprise at least one Albizia amaraseeds and at least one Magnolia officinalis barks.

A contemplated embodiment includes a preservative mixture or compositionthat comprises a mixture of at least one Albizia extract and at leastone Magnolia extract. Yet another contemplated embodiment includes acomposition that comprises a mixture of at least one Albizia extractenriched for one or more macrocyclic alkaloids and at least one Magnoliaextract enriched for one or more lignans. In contemplated embodiments,compositions are considered preservatives, antimicrobial agents orcombinations thereof.

Some contemplated embodiments comprise a mixture of at least one Albiziaextract from a plant material or other source and at least one Magnoliaextract from a plant material or another source, wherein the at leastone Albizia extract has an increase in an amount or an activity of oneor more macrocyclic alkaloids as compared to the amount or the activityof the one or more macrocyclic alkaloids in the plant material oranother source of the at least one Albizia extract, and wherein the atleast one Magnolia extract has an increase in an amount or an activityof one or more lignans as compared to the amount or the activity of theone or more lignans in the plant material or another source of the atleast one Magnolia extract.

In some embodiments, the at least one Albizia extract and the at leastone Magnolia extract are blended in a weight ratio ranging from about10:1 to 1:10. In other embodiments, the at least one Albizia extract andthe at least one Magnolia extract are blended in a weight ratio at about1:1. Contemplated preservatives, compositions and mixtures can beutilized to preserve food, cosmetics, nutraceuticals andpharmaceuticals. These contemplated preservatives, compositions andmixtures can be provided in a range from about 0.001% to about 10% byweight of the total composition. In some embodiments, the range is fromabout 0.01% to about 5% by weight of the total composition. In otherembodiments, the range is from about 0.1% to about 1% by weight of thetotal composition.

In addition, compositions, compounds and methods for preservatives andantimicrobial compositions are disclosed that include a mixture of plantextracts, wherein the plant extracts comprise at least one an Albiziaamara seeds extract enriched for one or more macrocyclic alkaloids and aMagnolia bark extract enrich for one or more biphenolic lignans.

The concept of discovering a unique blend of compounds and extracts withenhanced antimicrobial activities was borrowed from the practice ofcombinations of antibiotics. The field of synergistic combinations ofantibiotics is extremely broad and some fixed combinations weresuccessfully developed commercially, with respect to their effects ontreating infection with drug resistance. Through a high-throughputanti-E. coli screening of a plant library of over 10,216 organic extractand aqueous extracts, which are generated from medicinal plantscollected from all over the world, Albizia amara seeds extract wasidentified as the best lead exhibiting potent anti-Gram negativeinhibition with a MIC value of 40 ug/mL. Bioassay-guided isolation andidentification confirmed the active components are budmunchimaine C andits analogues.

The Albizia amara seeds extract showed potent inhibition against bothanti-Gram positive and anti-Gram-negative bacterial, but with mildanti-fungal activity against either Candida albicans or Aspergillusbrasiliensis. To search for a composition effective againstGram-positive, Gram-negative and fungi required by category 2 preservein USP chapter <51>, mainly targeting on boosting the anti-fungalactivity, specifically against C. albicans and A. brasiliensis, theAlbizia amara seeds extract was formulated with Magnolia barks extractfor unexpected enhanced outcomes.

Surprisingly and unexpectedly, the additions of Magnolia bark extractcaused a dramatic increase of its anti-fungal activity against both C.albicans and A. brasiliensis, and hence signify the importance of thecombination of these two materials. The merit of combining these twoplant materials were determined, and an unexpected synergy against bothC. albicans and A. brasiliensis was observed with the beneficial effectsexceeding the additive interaction based on the FCI index analysis.Collectively, it is contended that putting these two plants into thespecific ratios provides a novelty to the composition as demonstrated inits remarkable antibacterial activity and effects.

An interesting synergy was discovered when Albizia amara seeds extractwas blended with Magnolia bark extract at ratios of 1:3 and 1:1. Thesetwo plants were never combined together before at specific ratios toyield contemplated and disclosed bioactivities, which are generallyunderstood as the unique combination in these contemplated embodiments.

The antibacterial activity and efficacy of Composition 1A:1M, where Astands for Albizia amara seeds extract and M for Magnolia barks extract,was evaluated following the USP <51> guideline. Composition 1A:1M, whichis 1:1 ratio of Albizia amara seeds extract and Magnolia bark extract ata concentration of 1 mg/mL, significantly reduce the counts of P.aeruginosa, and A. brasiliensis with Log10 reduction at 5.58 and 4.1respectively and significantly, if not completely, eliminate S. aureus,E. coli, and C. albicans after 48 h, and no bacterial or fungi growthwas observed after 7 days, 14 days, and 28 days against all fivemicroorganisms. In fact, none of the constituents showed such potent andbroad antimicrobial activity at the magnitude equivalent to the oneshown for a contemplated compound or composition comprising Albiziaamara and Magnolia. Furthermore, a contemplated and unique ratio of orabout 1A:1M provides demonstrated effectiveness of this compositionagainst all five microorganisms including E. coli, P. aeruginosa, S.aureus, A. brasiliensis, and C. albicans.

It is contemplated that the Albizia is derived, obtained or selectedfrom at least one of the following—alone or in combination with oneanother: Albizia amara, Albizia schimperiana, Albizia lebbek, Albiziagummifera, Albizzia inopinata, Albizia saman (Samaneasaman=Pithecolobium saman), Albizia adianthifolia, Albizia adinocephala,Albizia anthelmintica, Albizia antunesiana, Albizia chevalieri, Albiziacorniculata, Albizia crassiramea, Albizia duclouxii, Albizia glabra,Albizia julibrissin, Albizia kalkora, Albizia lebbekoides, Albiziamyriophylla, Albizia nigricans, Albizia odoratissima, Albiziapeterisana, Albizia poilanei, Albizia procera, Albizia tanganyicensis,Albizia vialeana Pierre, Albizia zygia, Aphelandra fuscopunctata,Ephedra spp., Verbascum pseudonobile, Incarvillea sinensis, Verbascumphoenicum, Verbascum nigrum, Clerodendrum buchneri, or Aphelandrasquarrosa.

Albizia amara extract, as disclosed, is a contemplated component orconstituent that can be utilized as part of a target compound orcomposition. Albizia amara extract may be obtained, derived or selectedfrom any suitable source or sources, including Albizia schimperiana,Albizia lebbek, Albizia gummifera, Albizzia inopinata, Albizia saman(Samanea saman=Pithecolobium saman), Albizia adianthifolia, Albiziaadinocephala, Albizia anthelmintica, Albizia antunesiana, Albiziachevalieri, Albizia corniculata, Albizia crassiramea, Albizia duclouxii,Albizia glabra, Albizia julibrissin, Albizia kalkora, Albizialebbekoides, Albizia myriophylla, Albizia nigricans, Albiziaodoratissima, Albizia peterisana, Albizia poilanei, Albizia procera,Albizia tanganyicensis, Albizia vialeana Pierre, Albizia zygia,Aphelandra fuscopunctata, Ephedra spp., Verbascum pseudonobile,Incarvillea sinensis, Verbascum phoenicum, Verbascum nigrum,Clerodendrum buchneri, Aphelandra squarrosa or a combination thereof.

Albizia amara extract may be enriched for one or more macrocyclicalkaloids as contemplated herein. In some contemplated embodiments, theAlbizia extract comprises about 0.01% to 99.9% of macrocyclic alkaloids.Contemplated alkaloids isolated from Albizia or Albizia amara extractare extracted with any suitable solvent, including water, methanol,ethanol, alcohol, a water-mixed solvent or a combination thereof, orwith supercritical fluid. In contemplated embodiments, the Albizia amaraextract comprises about 0.01% to about 99.9% macrocyclic alkaloids.Contemplated alkaloids isolated from Albizia amara extract arebudmunchiamine A, B, C, D, E, F, G, H, I, J, K, budmunchiamine L1, L2,L3, L4, L5, L6, 13-normethylbudmunchiamine K, 9-normethylbudmunchiamineK, 1-normethylbudmunchiamine K, 6′-hydroxy-9-normethylbudmunchiamine K,6′-hydroxybudmunchiamine K, Felipealbizine A, Felipealbizine B,6′-hydroxybudmunchiamine C, Pithecolobine, Protomethine, Verbamethine,Verbametrine, Isoverbametrine, Isoverbamethine, Verbamekrine,Isoverbamekrine, Verbascine, Verbacine, Verbasitrine, Verballocine,Isoverbasitrine, Verballoscenine, Isoverbasikrine,N1-(Z)-p-Methoxycinnamoylbuchnerine, Buchnerine, Verbamedine,Verbascenine, Verbascenine, Isoverbamedine, Schweinine, Prelandrine,Incasine B′, Incasine B or a combination thereof.

Magnolia extract is a contemplated component or constituent that can beutilized as part of a target compound or composition. Magnolia extractmay be obtained from any suitable source, including Magnoliaofficinalis, Magnolia acuminate, Magnolia biondii, Magnolia coco,Magnolia denudate, Magnolia fargesii, Magnolia garrettii, Magnoliagrandiflora, Magnolia henryi, Magnolia liliflora, Magnoliakachirachirai, Magnolia Kobus, Magnolia obovata, Magnolia praecocissima,Magnolia pterocarpa, Magnolia pyramidata, Magnolia rostrate, Magnoliasalicifolia, Magnolia sieboldii, Magnolia soulangeana, Magnoliastellate, Magnolia virginiana, prod. of degradation of birch lignin,Acanthus ebracteatus, Aptosimum spinescens, Aralia bipinnata, Araucariaangustifolia, Araucaria araucana, Artemisia absinthium, Haplophyllumacutifolium, Haplophyllum perforatum, Liriodendron tulipifera, Krameriacystisoides, Perilla frutescens, Lawsonia inermis Myristica fragrans(nutmeg), Parakmeria yunnanensis (preferred genus name Magnolia), Perseajaponica, Piper futokadsura, Piper wightii, Rollinia mucosa, Sassafrasrandaiense, Scrophularia albida-colchica, Stellera chamaejasme, Syringavelutina, Syzygium cumini, Talauma gloriensis, Virola elongate,Urbanodendron verrucosum, Wikstroemia sikokiana or a combinationthereof.

Magnolia extract may be enriched for one or more lignans, includingdiphenol lignans, as contemplated herein. Contemplated lignans isolatedfrom Magnolia extract are extracted with any suitable solvent, includingwater, methanol, ethanol, alcohol, a water-mixed solvent or acombination thereof or with supercritical fluid.

In contemplated embodiments, the Magnolia extract comprises about 0.01%to about 99.9% diphenol lignans, biphenolic lignans or a combinationthereof. Contemplated lignans isolated from Magnolia extract aremagnolol, honokiol, Biondinin A,2,3-Bis(4-hydroxy-3-methoxyphenyl)-2-propenal, Denudadione C,Clovanemagnolol,8-[2-Hydroxy-5-(2-propenyl)phenyl]-6-(2-propenyl)-2H-1-benzopyran-2-one,Denudanolide C, Denudanolide D, Denudanolide B, Denudanolide A,Denudatin A, Denudatin B; (+)-form,4-Hydroxy-6′-methoxy-3,3′-biligna-7,7′-diene-9,9′-dial,4′-Methoxymagndialdehyde, 5,5′-Di-2-propenyl-[1,1′-biphenyl]-2,2′-diol,2,2′-Bichavicol, FEMA 4559, 5,5′-Diallyl-2,2′-dihydroxybiphenyl,Bornylmagnolol, 5,5′-Diallyl-2-(3-methyl-2-butenyloxy)biphenyl-2′-ol,5,5′-Diallyl-2-(allyloxy)biphenyl-2′-ol,3′,5-Diallyl-2,4′-dihydroxybiphenyl,3′,5-Di-2-propenyl-2,4′-biphenyldiol,5-Allyl-3′-(1-propenyl)biphenyl-2,4′-diol,3′-(1-Propenyl)-5-(2-propenyl)-2,4′-biphenyldiol,3′,5-Diallyl-2-hydroxy-4′-methoxybiphenyl, 4′-O-Methylhonokiol,2-Hydroxy-4′-methoxy-3′,5-di-2-propenylbiphenyl,4,4′-Diallyl-2,3′-dihydroxydiphenyl ether,2-[3-Hydroxy-4-(2-propenyl)phenoxy]-5-(2-propenyl)phenol,2-Hydroxy-3-methoxy-4′,5-di-2-propenyldiphenyl ether,4′,5-Diallyl-2-hydroxy-3-methoxydiphenyl ether,2-Methoxy-4-(2-propenyl)-6-[4-(2-propenyl)phenoxy]phenol, Fargesin,Methyl pluviatilol,3,4-Dimethoxy-3′,4′-methylenedioxy-7,9′:7′,9-diepoxylignan, Kobusin,Desmethoxyaschantin, O-Methylpiperitol, Spinescin, (+)-Fargesin,Planinin, Methylpluviatilol, Parakmerin A,6-Allyl-7-(3,4-dimethoxyphenyl)-2,3-dimethoxy-8-methyltricyclo[4.2.0.0.^(2,8)]oct-3-en-5-one,Magnosalin, Dipiperitylmagnolol, Kachirachirol B, Acuminatin, Licarin D,Eudeshonokiol A, Eudesmagnolol, Eudesmin; (+)-formm, (+)-Epieudesmin,(−)-Epieudesmin, Kachirachirol A, Eupomatenoid 13, Fargesiphenol B,Fargesiphenol C, Fargesone A, Fargesone B,4,5-Dimethoxy-3′,4′-methylenedioxy-2,7′-dioxo-5,8′-ligna-3,6,8-triene,4-(1,3-Benzodioxol-5-yl)-2,3,4,5-tetrahydro-7-methoxy-5-methyl-8H-2,5a-methano-1-benzoxepin-8-one,Garrettilignan A, Garrettilignan B,Tetrahydro-4-(3-hydroxy-4,5-dimethoxyphenyl)-1H,3H-furo[3,4-c]furan-1-one, Kachiranol,7′,9-Dihydroxy-3,3′,4,4′,5-pentamethoxy-7,9′-epoxylignan, Magnone B,3,3′,4,4′,5,9-Hexamethoxy-7,9′-epoxylignan-7′-one, Magnolenin C,3′,4,7,9,9′-Pentahydroxy-3,5′-dimethoxy-8,4′-oxyneolignan,3,3′-Dimethoxy-4,5:4′,5′-bis(methylenedioxy)pyramidatin,3′-Hydroxy-3,4,4′,5,5′-pentamethoxypyramidatin, Pyramidatin B,3,3′,4′,5′-Tetramethoxy-4,5-methylenedioxypyramidatin,3,3′,4,4′,5,5′-Hexamethoxypyramidatin, Pyramidatin C, Pyramidatin,Pyramidatin D, 3-Hydroxy-3′,4,4′,5,5′-pentamethoxypyramidatin,Pyramidatin A,3-Hydroxy-3′,4,5-trimethoxy-4′,5′-methylenedioxypyramidatin, PyramidatinF, Pyramidatin G, Pyramidatin H, Pyramidatin E,5′-Hydroxy-3,3′,4,4′,5-pentamethoxypyramidatin, Magnoshinin,2′,3,4,4′,5,6-Hexamethoxy-2,7′-cyclolign-7-ene, Acanthoside B,Eleutheroside E1,3-Hydroxy-3′,4,4′,5,5′-pentamethoxy-7,9′:7′,9-diepoxylignan,(+)-Episyringaresinol, Lirioresinol A, Symplicosigenol, Kobusinol B,Fargesol, 9-O-Acetylfargesol, Magnolone, Biondinin B, Biondinin E,Magnone A, 7,9,9′-Trihydroxy-3,3′,4-trimethoxy-8,4′-oxyneolignan,erythro-form, threo-form, Magnovatin A, Magnovatin B, Oleiferin C,3,4:3′,4′-Bis(methylendioxy)-7-lignanol,7-Hydroxy-3,4:3′,4′-bis(methylenedioxy)lignan, Liliflodione, DenudadioneB, Denudadione A, Liliflol A, Liliflol B, Denudatone,3-[2′,6-Dihydroxy-5′-(2-propenyl)[1,1′-biphenyl-3-yl]]-2-propenal,Randainal, Magnaldehyde B, Magnaldehyde C, 6′-O-Methylmagnaldehyde B,4-Deoxy-6-methoxymagnaldehyde A, 4-Methoxymagnaldehyde B,2′,6-Dihydroxy-5′-(2-propenyl) [1,1′-biphenyl]-3-carboxaldehyde,5-Allyl-5′-formyl-2,2′-dihydroxybiphenyl, Magnaldehyde E,6-O-Methylmagnaldehyde E,9-Hydroxy-3,3′,4,4′,5-pentamethoxy-7,9′-epoxylign-7′-ene,9,9′-Dihydroxy-3,3′,4,4′,5-pentamethoxy-2,7′-cyclolign-7-ene,Magnolianin, Magnolignan I, Magnolignan B, Magnolignan A, Magnolignan D,Magnolignan C 6′-glucoside, Magnolignan C, Magnolignan E, Magnolignan F,Magnolignan G, Magnolignan H,2-[4-Hydroxy-3-(2-propenyl)phenyl]-2,5-cyclohexadiene-1,4-dione,2-(3-Allyl-4-hydroxyphenyl)-1,4-benzoquinone, Magnosalicin;(7RS,7′SR,8RS,8′RS)-form, Magnostellin B, Magnostellin B; 7,9-Diepimer,5-methoxy,5-[2-Hydroxy-5-(2-propenyl)phenyl]-2-methylbenzofuran,3-Hydroxy-3′,4,4′,5-tetramethoxy-7,9′,7′,9-diepoxylignan,3-De-O-methylmagnolin, Magnolin, 3-O-Demethylaschantin,3′-O-Demethylaschantin,3-Hydroxy-4,5-dimethoxy-3′,4′-methylenedioxy-7,9′:7′,9-diepoxylignan,5′-Hydroxy-4′-O-methylpiperitol, Epimagnolin A,7′-Hydroxy-3,3′,4,4′-tetramethoxy-7,9′-epoxylignan, Magnostellin A,Kobusinol A, 4,7′-Dihydroxy-3,3′,4′-trimethoxy-7,9′-epoxylignan,4′,5,9-Trihydroxy-3,3′-dimethoxy-7,9′-epoxylignan, Fargesiphenol A,Liliflone, Monoterpenylmagnolol, Piperitylhonokiol,Monoterpenylhonokiol, Saulangianin I, Sesamin; (−)-form, Syringinoside,Syringin 4″-glucoside, 3,3′,4,5′-Tetramethoxy-7,9′:7′,9-diepoxylignan,3′,5′-Dimethoxy-3,4-methylenedioxy-7,9′:7′,9-diepoxylignan, Galgravin,2,2′-Dihydroxy-3-methoxy-5,5′-di-2-propenylbiphenyl, 3-Methoxymagnolol,5,5′-Diallyl-2,2′-dihydroxy-3-methoxybiphenyl or a combination of any ofthese thereof.

Contemplated compounds, medicinal compositions and compositions maycomprise or additionally comprise or consist of at least oneantimicrobial ingredient. In some embodiments, the at least oneantimicrobial ingredient may comprise or consist of plant powder orplant extract of Psoralea corylifolia, Curcumin, Curcuma longa, Eugenol,Alpinia galganga, coptidis, Azadirachta indica, Propolis, clove oil,Sophora flavescens, Areca catechu, Glycyrrhiza glabra, Thuja plicata,Cortex phellodendri, Rheum palate, Alpinia officinarum, Broussonetiapapyriferra, Viburnum cotinifolium, Euphorbia hirta Linn, Vitex negundo,or a combination thereof.

Contemplated compounds, medicinal compositions and compositions maycomprise or additionally comprise, consist of or consist essentially ofat least one or more known antimicrobial preservatives, constituents oringredients. In some embodiments, the at least one antimicrobialingredient may comprise or consist of calcium propionate, sodiumbenzoate, sodium nitrate, sodium nitrite, potassium sorbate, benzoicacid, salicylic acid, sorbic acid, alcohol, penoxyethanol, 1,2-diolalcohol (1,2 propanediol, 1,2 butanediol, 1,2 pentanediol,1,2-hexanediol), 1,3-diol alcohol (1,3 propanediol, 1,3 butanediol, 1,3pentaneidol, 1,3-hexanediol), 1,5 pentanediol, benzakonium chloride,parabens, sulfites (sulfur dioxide, sodium bisulfite, potassium hydrogensulfite, etc.) and disodium EDTA, and BHT (butylated hydroxy toluene)and BHA (butylated hydroxyl).anisole), formaldehyde (usually insolution), glutaraldehyde (kills insects), andmethylchloroisothiazolinone.

The composition further comprises a pharmaceutically or nutraceuticallyor cosmetically acceptable carrier, diluent, or excipient, wherein thepharmaceutical or nutraceutical formulation comprises from about 0.001weight percent (wt %) to about 10 wt % of the active ingredients of theextract mixture. These contemplated active ingredients can be providedin a range from about 0.001% to about 10% by weight of the totalcomposition. In some embodiments, the range is from about 0.01% to about5% by weight of the total composition. In other embodiments, the rangeis from about 0.1% to about 1% by weight of the total composition.

Also, contemplated herein are preservatives or antimicrobial agents ofthe disclosed compounds. Such products may result from, for example, theoxidation, reduction, hydrolysis, amidation, esterification, and thelike of the administered compound, primarily due to enzymatic processes.Accordingly, contemplated compounds are those produced by a processcomprising administering a contemplated compound or composition to amammal for a period of time sufficient to yield a metabolic productthereof. Such products are typically identified by administering aradiolabeled compound of this disclosure in a detectable dose to ananimal, such as rat, mouse, guinea pig, dog, cat, pig, sheep, horse,monkey, or human, allowing sufficient time for metabolism to occur, andthen isolating its conversion products from the urine, blood or otherbiological samples.

As used herein, the phrases “stable compound” and “stable structure” areused interchangeably and used to indicate a compound that issufficiently robust to survive isolation to a useful degree of purityfrom a reaction mixture, and to survive formulation into an efficacioustherapeutic agent.

As used herein, the term “mammal” includes humans and both domesticanimals, such as laboratory animals or household pets (e.g., rat, mouse,guinea pig, cats, dogs, swine, cattle, sheep, goats, horses, rabbits,primates), and non-domestic animals, such as wildlife or the like.

As used herein, the terms “optional” or “optionally” may be usedinterchangeably and mean that the subsequently described element,component, event or circumstances may or may not occur, and includesinstances where the element, component, event or circumstance occur andinstances in which they do not. For example, “optionally substitutedaryl” means that the aryl radical may or may not be substituted—in otherwords, the description includes both substituted aryl radicals and arylradicals having no substitution.

Contemplated compounds, medicinal compositions and compositions maycomprise or additionally comprise or consist of at least onepharmaceutically or nutraceutically or cosmetically acceptable carrier,diluent or excipient. As used herein, the phrase “pharmaceutically ornutraceutically or cosmetically acceptable carrier, diluent orexcipient” includes any adjuvant, carrier, excipient, glidant,sweetening agent, diluent, preservative, dye/colorant, flavor enhancer,surfactant, wetting agent, dispersing agent, suspending agent,stabilizer, isotonic agent, solvent, or emulsifier which has beenapproved by the United States Food and Drug Administration as beingacceptable for use in humans or domestic animals.

Contemplated compounds, medicinal compositions and compositions maycomprise or additionally comprise or consist of at least onepharmaceutically or nutraceutically or cosmetically acceptable salt. Asused herein, the phrase “pharmaceutically or nutraceutically orcosmetically acceptable salt” includes both acid addition and baseaddition salts.

As used herein, the phrase “pharmaceutically or nutraceutically orcosmetically acceptable acid addition salt” refers to those salts whichretain the biological effectiveness and properties of the free bases,which are not biologically or otherwise undesirable, and which areformed with inorganic acids such as hydrochloric acid, hydrobromic acid,sulfuric acid, nitric acid, phosphoric acid and the like, and organicacids such as acetic acid, 2,2-dichloroacetic acid, adipic acid, alginicacid, ascorbic acid, aspartic acid, benzenesulfonic acid, benzoic acid,4-acetamidobenzoic acid, camphoric acid, camphor-10-sulfonic acid,capric acid, caproic acid, caprylic acid, carbonic acid, cinnamic acid,citric acid, cyclamic acid, dodecylsulfuric acid, ethane-1,2-disulfonicacid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, formic acid,fumaric acid, galactaric acid, gentisic acid, glucoheptonic acid,gluconic acid, glucuronic acid, glutamic acid, glutaric acid,2-oxo-glutaric acid, glycerophosphoric acid, glycolic acid, hippuricacid, isobutyric acid, lactic acid, lactobionic acid, lauric acid,maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonicacid, mucic acid, naphthalene-1,5-disulfonic acid,naphthalene-2-sulfonic acid, 1-hydroxy-2-naphthoic acid, nicotinic acid,oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid,propionic acid, pyroglutamic acid, pyruvic acid, salicylic acid,4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid,tartaric acid, thiocyanic acid, p-toluenesulfonic acid, trifluoroaceticacid, undecylenic acid, or the like.

As used herein, the phrase “pharmaceutically or nutraceutically orcosmetically acceptable base addition salt” refers to those salts whichretain the biological effectiveness and properties of the free acids,which are not biologically or otherwise undesirable. These salts areprepared from addition of an inorganic base or an organic base to thefree acid. Salts derived from inorganic bases include the sodium,potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper,manganese, aluminum salts and the like. In certain embodiments, theinorganic salts are ammonium, sodium, potassium, calcium, or magnesiumsalts. Salts derived from organic bases include salts of primary,secondary, and tertiary amines, substituted amines including naturallyoccurring substituted amines, cyclic amines and basic ion exchangeresins, such as ammonia, isopropylamine, trimethylamine, diethylamine,triethylamine, tripropylamine, diethanolamine, ethanolamine, deanol,2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine,lysine, arginine, histidine, procaine, hydrabamine, choline, betaine,benethamine, benzathine, ethylenediamine, glucosamine, methylglucamine,theobromine, triethanolamine, tromethamine, purines, piperazine,piperidine, N-ethylpiperidine, polyamine resins and the like.Particularly useful organic bases include isopropylamine, diethylamine,ethanolamine, trimethylamine, dicyclohexylamine, choline, or caffeine.

Often crystallizations produce a solvate of or include contemplatedcompounds. As used herein, the term “solvate” refers to an aggregatethat comprises one or more molecules of a contemplated compound,medicinal composition or composition with one or more molecules ofsolvent. The solvent may be water, in which case the solvate may be ahydrate. Alternatively, the solvent may be an organic solvent. Thus, thecontemplated compounds, medicinal compositions or compositions may existas a hydrate, including a monohydrate, dihydrate, hemihydrate,sesquihydrate, trihydrate, tetrahydrate and the like, as well as thecorresponding solvated forms. A contemplated compound, medicinalcomposition or composition may be a true solvate, while in other cases,a contemplated compound, medicinal composition or composition may merelyretain adventitious water or be a mixture of water plus someadventitious solvent.

A preservative or antimicrobial agent refers to a formulation of acontemplated compound, composition or medicinal composition and a mediumgenerally accepted in the art for the delivery of the antimicrobialactivity in food, cosmetics, nutraceuticals, oral hygiene products, andpharmaceuticals. For example, a contemplated preservative compound, orcomposition may be formulated or used as a stand-alone composition, oras a component in a prescription drug, an over-the-counter (OTC)medicine, a botanical drug, an herbal medicine, a homeopathic agent, orany other form of health care product reviewed and approved by agovernment agency. Exemplary and contemplated preservative compositionsmay be formulated or used as a stand-alone composition, or as anutritional or bioactive component in food, a novel food, a functionalfood, a beverage, a bar, a food flavor, a food additive, a medical food,a dietary supplement, or an herbal product. A medium generally acceptedin the art includes all pharmaceutically or nutraceutically acceptablecarriers, diluents or excipients therefor.

In some embodiments, contemplated compositions are formulated as atablet, hard capsule, soft gel capsule, powder, granule, liquid,tincture, sashay, ready to drink shot, or lozenge, cosmetic cream, gel,lotion, shampoo, hire conditioner, body rinse, body lotion, hire coloragent, skin color agent, eye color agent, lip burn, perfume, toothpaste,mouth washing, fruit juice, aloe juice, yogurt, foods, soft drink,beverage, processed fruits, cheese products, wine, gem, peanut butter,RTD, protein bar, or a snack bar.

As used herein, the phrase “enriched for” refers to a plant extract orother preparation having at least about a two-fold up to about a1000-fold increase in the amount or activity of one or more activecompounds as compared to the amount or activity of the one or moreactive compounds found in the weight of the plant material or othersource before extraction or other preparation. In certain embodiments,the weight of the plant material or other source before extraction orother preparation may be dry weight, wet weight, or a combinationthereof.

As used herein, “major active ingredient” or “major active component”refers to one or more active contemplated compounds found in a plantextract or other preparation, or enriched for in a plant extract orother preparation, which is capable of at least one biological activity.In certain embodiments, a major active ingredient of an enriched extractwill be the one or more active compounds that were enriched in thatextract. Generally, one or more major active components will impart,directly or indirectly, most (i.e., greater than 50%) of one or moremeasurable biological activities or effects as compared to other extractcomponents. In certain embodiments, a major active ingredient may be aminor component by weight percentage of an extract (e.g., less thanabout 50%, 25%, 20%, 15%, 10%, 5%, or 1% of the components contained inan extract) but still provide most of the desired biological activity.Any contemplated composition containing a major active ingredient mayalso contain minor active ingredients that may or may not contribute tothe pharmaceutical or nutraceutical activity of the enrichedcomposition, but not to the level of major active components, and minoractive components alone may not be effective in the absence of a majoractive ingredient.

As used herein, the phrases “effective amount” or “therapeuticallyeffective amount” refer to that amount of a contemplated compound,medicinal composition or composition that, when used in food, cosmetics,nutraceuticals, oral hygiene products, and pharmaceuticals, issufficient to prevent the degradation of bacterial and inhibit thegrowth of bacteria, fungi and other microorganisms.

The chemical naming protocol and any structure diagrams used herein area modified form of the I.U.P.A.C. nomenclature system, using theACD/Name Version 9.07 software program or ChemDraw Ultra Version 11.0software naming program (CambridgeSoft), wherein the compounds of thisdisclosure are named herein as derivatives of the central corestructure, e.g., the imidazopyridine structure. For complex chemicalnames utilized herein, a substituent group is named before the group towhich it attaches. For example, cyclopropylethyl comprises an ethylbackbone with cyclopropyl substituent.

A contemplated preservative composition may be in the form of a solid orliquid. In one aspect, the carrier(s) are particulate, so that thecompositions are, for example, in tablet or powder form. The carrier(s)may be liquid, with the compositions being, for example, oral syrup,injectable liquid or an aerosol, which is useful in, for example,inhalatory administration.

When intended for oral administration, the pharmaceutical ornutraceutical composition is in either solid or liquid form, where semisolid, semi liquid, suspension and gel forms are included within theforms considered herein as either solid or liquid.

As a solid composition for oral administration, the preservativecomposition may be formulated into a powder, granule, compressed tablet,pill, capsule, chewing gum, wafer, bar, or like form. Such a solidcomposition will typically contain one or more inert diluents or ediblecarriers. In addition, one or more of the following may be present:binders such as carboxymethylcellulose, ethyl cellulose, cyclodextrin,microcrystalline cellulose, gum tragacanth or gelatin; excipients suchas starch, lactose or dextrins, disintegrating agents such as alginicacid, sodium alginate, Primojel®, corn starch and the like; lubricantssuch as magnesium stearate or Sterotex®; glidants such as colloidalsilicon dioxide; sweetening agents such as sucrose or saccharin; aflavoring agent such as peppermint, methyl salicylate or orangeflavoring; and a coloring agent.

A contemplated preservative composition may be in the form of a capsule,for example, a gelatin capsule, it may contain, in addition to materialsof the above type, a liquid carrier such as polyethylene glycol or oil.

A contemplated preservative composition may be in the form of a liquid,for example, an elixir, syrup, gel, solution, emulsion or suspension.The liquid may be for oral administration or for delivery by injection,as two examples. When intended for oral administration, a usefulcomposition contains, in addition to the present compounds, one or moreof a sweetening agent, preservatives, dye/colorant and flavor enhancer.In a composition intended to be administered by injection, one or moreof a surfactant, preservative, wetting agent, dispersing agent,suspending agent, buffer, stabilizer and isotonic agent may be included.

A contemplated liquid preservative compositions, whether they besolutions, suspensions or other like form, may include one or more ofthe following adjuvants: sterile diluents such as water for injection,saline solution, such as physiological saline, Ringer's solution,isotonic sodium chloride, fixed oils such as synthetic mono ordiglycerides which may serve as the solvent or suspending medium,polyethylene glycols, glycerin, propylene glycol or other solvents;antibacterial agents such as benzyl alcohol or methyl paraben;antioxidants such as ascorbic acid or sodium bisulfite; chelating agentssuch as ethylenediaminetetraacetic acid; buffers such as acetates,citrates or phosphates and agents for the adjustment of tonicity such assodium chloride or dextrose. The parenteral preparation can be enclosedin ampoules, disposable syringes or multiple dose vials made of glass orplastic. Physiological saline is a generally useful adjuvant. Aninjectable pharmaceutical or nutraceutical composition is sterile.

A contemplated liquid preservative composition intended for eitherparenteral or oral administration should contain an amount of acontemplated compound, medicinal composition or composition such that asuitable dosage will be obtained.

A contemplated preservative composition may be intended for topicaladministration, in which case the carrier may suitably comprise asolution, emulsion, cream, lotion, ointment, or gel base. The base, forexample, may comprise one or more of the following: petrolatum, lanolin,polyethylene glycols, bee wax, mineral oil, diluents such as water andalcohol, and emulsifiers and stabilizers. Thickening agents may bepresent in a pharmaceutical or nutraceutical composition for topicaladministration. If intended for transdermal administration, thecomposition may include a transdermal patch or iontophoresis device.

A contemplated preservative composition may be intended for rectaladministration, in the form, for example, of a suppository, which willmelt in the rectum and release the drug. The composition for rectaladministration may contain an oleaginous base as a suitablenonirritating excipient. Such bases include lanolin, cocoa butter andpolyethylene glycol.

A contemplated preservative composition may include various materials,which modify the physical form of a solid or liquid dosage unit. Forexample, the composition may include materials that form a coating shellaround the active ingredients. The materials that form the coating shellare typically inert, and may be selected from, for example, sugar,shellac, and other enteric coating agents. Alternatively, the activeingredients may be encased in a gelatin capsule.

A contemplated preservative composition in solid or liquid form mayinclude an agent that binds to the contemplated compound and therebyassists in the delivery of the compound. Suitable agents that may act inthis capacity include a monoclonal or polyclonal antibody, a protein ora liposome.

A contemplated preservative composition in solid or liquid form mayinclude reducing the size of a particle to, for example, improvebioavailability. The size of a powder, granule, particle, microsphere,or the like in a composition, with or without an excipient, can be macro(e.g., visible to the eye or at least 100 μm in size), micro (e.g., mayrange from about 100 μm to about 100 nm in size), nano (e.g., may nomore than 100 nm in size), and any size in between or any combinationthereof to improve size and bulk density.

A contemplated preservative composition may be prepared by methodologywell known in the preservative art. For example, a preservativecomposition intended to be administered by injection can be prepared bycombining a contemplated compound with sterile, distilled water so as toform a solution. A surfactant may be added to facilitate the formationof a homogeneous solution or suspension. Surfactants are compounds thatnon-covalently interact with a contemplated compound so as to facilitatedissolution or homogeneous suspension of the compound in the aqueousdelivery system.

Contemplated compounds, compositions and medicinal compositions, ortheir pharmaceutically or nutraceutically acceptable salts, areadministered in a therapeutically effective amount, which will varydepending upon a variety of factors including the activity of thespecific compound employed; the metabolic stability and length of actionof the compound; the age, body weight, general health, sex, and diet ofthe patient; the mode and time of administration; the rate of excretion;the drug combination; the severity of the particular disorder orcondition; and the subject undergoing therapy.

Contemplated compounds, compositions and medicinal compositions, orpharmaceutically or nutraceutically acceptable derivatives thereof, mayalso be administered simultaneously with, prior to, or afteradministration of one or more other therapeutic agents. Such combinationtherapy includes administration of a single pharmaceutical ornutraceutical dosage formulation that contains a contemplated compoundand one or more additional active agents, as well as administration of acontemplated compound and each active agent in its own separatepharmaceutical or nutraceutical dosage formulation. For example, acontemplated compound and another active agent can be administered tothe patient together in a single oral dosage composition, such as atablet or capsule, or each agent can be administered in separate oraldosage formulations. Where separate dosage formulations are used,contemplated compounds and one or more additional active agents can beadministered at essentially the same time, i.e., concurrently, or atseparate staggered times, i.e., sequentially; combination therapy isunderstood to include all these regimens.

It is understood that in the present description, combinations ofsubstituents or variables of the depicted formulae are permissible onlyif such contributions result in stable compounds.

It will also be appreciated by those skilled in the art that in theprocess described herein the functional groups of intermediate compoundsmay need to be protected by suitable protecting groups. Such functionalgroups include hydroxy, amino, mercapto and carboxylic acid. Suitableprotecting groups for hydroxy include trialkylsilyl or diarylalkylsilyl(for example, t-butyldimethylsilyl, t-butyldiphenylsilyl ortrimethylsilyl), tetrahydropyranyl, benzyl, and the like. Suitableprotecting groups for amino, amidino and guanidino includet-butoxycarbonyl, benzyloxycarbonyl, and the like. Suitable protectinggroups for mercapto include C(O)R″ (where R″ is alkyl, aryl orarylalkyl), p-methoxybenzyl, trityl and the like. Suitable protectinggroups for carboxylic acid include alkyl, aryl or arylalkyl esters.Protecting groups may be added or removed in accordance with standardtechniques, which are known to one skilled in the art and as describedherein. The use of protecting groups is described in detail in Green, T.W. and P. G. M. Wutz, Protective Groups in Organic Synthesis (1999), 3rdEd., Wiley, which is incorporated by reference herein in its entirety.As one of skill in the art would appreciate, a protecting group may alsobe a polymer resin such as a Wang resin, Rink resin or a2-chlorotrityl-chloride resin.

It will also be appreciated by those skilled in the art, although suchprotected derivatives of contemplated compounds may not possesspharmacological activity as such, they may be administered to a mammaland thereafter metabolized in the body to form compounds which arepharmacologically active. Such derivatives may therefore be described as“prodrugs”. All prodrugs of contemplated compounds are included withinthe scope of this disclosure.

Furthermore, contemplated compounds that exist in free base or acid formcan be converted to their pharmaceutically or nutraceutically acceptablesalts by treatment with the appropriate inorganic or organic base oracid by methods known to one skilled in the art. Salts of contemplatedcompounds can be converted to their free base or acid form by standardtechniques.

In some embodiments, contemplated compounds, compositions and/ormedicinal compositions can be isolated from plant sources, for example,from those plants included in the examples and elsewhere throughout thepresent application. Suitable plant parts for isolation of contemplatedextracts and compounds include leaves, bark, trunk, trunk bark, stems,stem bark, twigs, tubers, root, root bark, bark surface (such asperiderm or polyderm, which may include phellem, phellogen, phelloderm,or any combination thereof), young shoots, rhizomes, seed, fruit,androecium, gynoecium, calyx, stamen, petal, sepal, carpel (pistil),flower, or any combination thereof. Contemplated plant extracts arederived from at least one plant part selected from the group consistingof stems, stem barks, trunks, trunk barks, twigs, tubers, roots, rootbarks, young shoots, seeds, rhizomes, flowers and other reproductiveorgans, tissue cultures and stem cell cultures, leaves, other aerialparts or a combination thereof. In some related embodiments,contemplated compounds are isolated from plant sources and syntheticallymodified to contain any of the recited substituents. In this regard,synthetic modification of contemplated compounds isolated from plantscan be accomplished using any number of techniques that are known in theart and are well within the knowledge of one of ordinary skill in theart.

EXAMPLES Example 1 Preparation of Organic Extracts

Dried ground plant materials (20 g) were loaded into two 100 mLstainless steel tubes and extracted twice with an organic solventmixture (methylene chloride/methanol in a ratio of 1:1) using an ASE 300automatic extractor at 80° C. and pressure 1500 psi. The extractsolution is automatically filtered and collected. The organic extractsolution is evaporated with rotary evaporator to give crude organicextract (OE). The stainless steel tube were flushed with fresh solventand purged with nitrogen gas to dryness before switching to aqueousextraction at 50° C. The aqueous solution was filtered and freeze-driedto provide aqueous extract (AE).

Example 2 High Throughput Screening for Anti-Microbial Activity AgainstEscherichia Coli

The plant library with 10, 216 OE and AE were screened for antimicrobialactivity against E. coli with a high throughput screening in 96-wellmicrotiter plates. Microbial cultures E. coli KCTC2571 (=ATCC8739) wasused as the test microorganisms. LB broth was used as the diluent ormedia in the wells. Cultures of bacteria were incubated for 24 h at 37°C. The plant library samples were screened in duplicate for each sampleat a concentration of 250 ug/mL. 180 ul of E. coli culture was added ineach well with inoculum size of 3×10⁵ CFU/mL and mixed with 20 ul testsample. 200 ul bacterial cultures without treatment were used as controlwith 200 ul LB broth as blank. Cultures of bacteria were incubated for24 h at 37° C. and OD was measured at 600 nm for each sample. Theinhibition was calculated based on the following equation. Grape seedsextract was used as positive control. The preliminary results showed 82extracts with over 80% inhibitions at 250 ug/mL, which were furthertested at 50 ug/mL. 11 plant extracts exhibited 80% or higher inhibitionat the concentration of 50 ug/mL. MIC values were determined for those11 hits, using the same method with serial dilutions. The plant hitswere selected for further development based on the MIC₉₉ values. Albiziaamara seed was the top hit selected from this screening with potentanti-E. coli activity with a MIC value at 12.5 ug/mL.

${{Inhibition}\mspace{14mu}(\%)} = {\frac{{Control} - \left( {{Sample} - {Blank}} \right)}{Control} \times 100}$

Example 3 Preparation of Organic Extracts from Albizia Amara Seeds

A total of 100 grams of dried Albizia amara seed powder are loaded intofive 100 mL stainless steel tube and extracted twice with methanol usingan ASE 350 automatic extractor at 80 degree and pressure 1500 psi. Theextract solution is automatically filtered and collected. The organicextract solution is evaporated with rotary evaporator to give crudeorganic extract (OE) (25.1 g, 25.1%).

Similar results were obtained using the same procedure, but with theorganic solvent being replaced with methanol or ethanol to provide amethanol extract (ME) or ethanol extract (EE), methanol:H₂O (7:3)extracts, methanol:H₂O (1:1) extracts, methanol:H₂O (3:7) ethanol:H₂O(7:3) extracts, ethanol:H₂O (1:1) extracts, ethanol:H₂O (3:7) extractsand water extracts respectively.

TABLE 1 The Albizia amara seed extracts extracted with differentsolvents Collect Extraction ID Species Parts Ext_ID Yield (%) SolventR00557 Albizia Seed RN368-28-01 27.6 Water amara RN368-29-01 24.5 MeOHRN368-29-02 26.5 50% MeOH/H2O

Example 4 Fractionation, Purification and Identification andQuantification of Active Compounds from Albizia Amara Seeds Extracts

Bioassay guided isolation lead to the isolation and identification ofthe active macrocyclic alkaloid budmunchiamine C. 2 g Albizia seedsmethanol extract was loaded to a C18 packed open column eluted with asolvent system of mixture of MeOH and 0.05% trifluoroacetic acid inwater in a range of 55% MeOH to 75% MeOH. 9 fractions were collected.The active fraction 40 mg was dissolved in water 4 mL. Then Ethylacetate 8 mL was added to the sample. After centrifuge, the lower layerwas collected and this procedure will be repeated for three times. Thenthe sample will be dissolved in ammonium hydroxide water 4 mL with a PHvalue of 11 and washed with ethyl acetate 8 mL for three times to givethe budmunchiamine C (23.5 mg). Methanol/ethanol was selected as thefinal solvent for extraction because methanol or ethanol extract showedhighest alkaloid contents in the final extraction.

Budmunchiamine C content in the extracts extract by different solventsystem could be analyzed by HPLC method with using a RP-C18 column(Phenomenex, 4.6×250 mm, 5 μm), with a gradient elution by methanol and0.5% TFA in Water at a flow rate of 1 mL/min detected by UV detector at210 nm or ELSD detector. Budmunchiamine C peak was identified based onthe comparison with standard compound isolated and identified in house.Methanol extract (RN368-29-01) has the highest budmunchiamine C contentcompared with water, and 50% Methanol/H2O extracts, and showed the bestanti-E. coli activity with a MIC value of 40 ug/mL. This result isconsistent with the discovery and identification budmunchiamine C andits analogues as the active components responsible for the anti-GramNegative bacterial activity of this plant. The budmunchiamines contentin the plant extracts from different species, parts and collected fromdifferent location and season could be varied in a range of 0-6%. Themethanol extract of the Albizia amara seeds was identified as the goodsource for budmunchiamine type alkaloids.

TABLE 2 Budmunchiamine C content and MIC against E. coli of the Albiziaamara seed extracts Extract ID Extraction Budmunchiamine C MIC (ug/mL)RN368-29-01 Methanol extract 1.8% 40 RN368-29-02 50% Methanol/ 1.6% 100water extract RN368-28-01 Water extract 1.1% 100

Example 5 Anti-Microbial Activities of Different Albizia Species

To search for the Albizia material with good anti-Gram negativebacterial activity, Albizia plants of different species and differentparts were collected from several countries including China, India,Pananma, Gana and Zimbabwe with diverse geological locations. Theanti-E. coli activity was evaluated and MIC values were determined forthe extracts of each Albizia material following the protocol as depictedin Example 2 with the anti-microbial activities results shown in Table3.

TABLE 3 in vitro Anti-E. coli activity of Albizia extracts Plant speciesCollect ID Plant parts Country origin MIC (ug/mL) A. amara R00557 seedsIndia 40 A. amara R00224 seeds Zimbabwe >100 A. julibrissin R00558 seedsChina >100 A. lebbeck R00584 seeds India >100 A. procera R00585 seedsIndia >100 A. richardiana R00600 seeds India >100 A. falcataria R00610seeds India >100 A. amara R00581 Leaf India >100 A. amara R00582 BarkIndia >100 A. amara R00583 Root India >100 A. petersana R00226 Root barkZimbabwe >100 A. gumifera R00225 Stem bark Zimbabwe >100 A. julibrissinR00554 Flower China >100 A. julibrissin R00555 Bark China >100 A.kalkora R00567 Bark China >100 A. lebbeck R00568 Bark China >100 A.lebbeck E1373 Bark India >100 A. amara R00581 Leaf India >100 A. amaraR00582 Bark India >100 A. amara R00583 Root India >100 A. seman P03417Leaf Panama 250 A. seman P01124 Leaf Ghana >150 A. seman P01125 Rootbark Ghana 150 A. seman P01126 Stem bark Ghana 150

Example 6 Antimicrobial Activity of Albizia Amara Seeds Extract andBudmunchiamine C

Minimum Inhibitory Concentration against bacterial S. aureus, P.aeruginosa, E. coli, and fungi A. brasiliensis and C. albicans weredetermined for Albizia amara seeds extract and the active markercompound bundmunchiamine C. The Minimum Inhibitory Concentration Method(MIC) is carried out following the general microbiological protocol tomeasure the lowest level of an antimicrobial agent that can inhibitmicrobial proliferation in liquid. Test microorganisms are prepared inliquid culture medium for bacteria or on agar for fungi. Suspensions oftest microorganisms are standardized by dilution in appropriate brothsolution, usually Mueller-Hinton broth. The test substance is preparedby conducting several serial 1:1 dilutions in a 96-well microtiter plateor in small test tubes, through Mueller-Hinton broth or otherappropriate medium. All wells or tubes containing diluted testsubstances are inoculated with test microorganisms, individually,resulting in one additional and final dilution of the product in alltest vessels. The microtiter plate or test tubes are incubated for 18-24hours. After the incubation period, observations are made to determinethe minimum concentration (MIC) which is the lowest concentration oftest substance in a well where no turbidity (indicative of testmicroorganism growth) is observed and minimum bactericidal concentration(MBC).

The Albizia amara seeds extract RN368-31-MIX and budmunchiamine C weretested against five microorganisms required for category 2 includingStaphylococcus aureus (ATCC 6538), Escherichia coli (ATCC 8739),Pseudomonas aeruginosa (ATCC 9027), Candida albicans (ATCC 10231),Aspergillus brasiliensis (ATCC 16404). Both Albizia amara seeds extractand budmunchiamine C showed potent anti-E. coli activity with MIC valuesat 62.5 ug/mL for the seed extract, and 31.25 for budmunchiamine C.Relatively weaker anti-fungal activities were observed for the seedextract and budmunchiamine C. Budmunchiamine C was further testedagainst six other microorganisms with both MIC and MBC results shown inTable 5. Budmunchiamine C showed the most potent efficacy againstStreptococcus pyogenes with a MIC of <0.97 ug/mL. It also showedsignificant efficacies against Trichophyton mentagrophytes, Trichophytonrubrum, Propionibacterium acnes, and Staphylococcus epidermis.

TABLE 4 Anti-microbial activities of Albizia amara seeds extractRN368-31-MIX and budmunchiamine C. Budmunchiamine C RN368-31-MIX MICOrganism MIC (ug/mL) (ug/mL) MBC (ug/mL) Staphylococcus aureus 125 31.2531.25 Escherichia coli 62.5 31.25 31.25 Pseudomonas aeruginosa 250 62.562.5 Candida albicans negative * 125 Aspergillus brasiliensis 500125 * * Unable to be determined

TABLE 5 Anti-microbial activities of budmunchiamine C against othermicroorganism Organism MIC MBC Microsporum canis >0.0375% (375ug/mL) >0.0375% (375 ug/mL) Trichophyton 0.001172% (11.7 ug/mL)0.001172% (11.7 ug/mL) mentagrophytes Trichophyton rubrum 0.001172%(11.7 ug/mL) 0.004688% (46.8 ug/mL) Propionibacterium acnes 0.0001953%(1.95 ug/mL) 0.00078125% (7.8 ug/mL) Staphylococcus 0.003125% (31.25ug/mL) 0.003125% (31.25 ug/mL) epidermis Streptococcus pyogenes<0.00009765% (<0.97 ug/mL) <0.000097656% (<0.97 ug/mL)

Example 7 Analytical Method for Analysis of Magnolia Stem Bark Extracts

The Magnolia stem bark extracts were analyzed by RP-HPLC method byquantification of active marker compounds honokiol and magnolol. Theextract was prepared as a methanol solution sonicated for approximately10 minutes. The flask was cooled to room temperature and QS withextraction method; mixed well and filtered through a 0.45 um nylonsyringe filter and 20 μl solutions were injected and analyzed, magnololand honokiol content in Magnolia stem bark extracts were quantified byRP-HPLC method on an Agilent HPLC/PDA system with the C18 column(Phenomenex, USA). A binary purified water and acetonitrile solventsystem was used for detection of magnolol and honokiol with isocraticelution of 77% acetonitrile in water for 18 min at a flow rate of 1mL/min with a column temperature of 35° C. at a wavelength of 290 nm.The honokiol and magnolol content in the Magnolia stem bark Extract(FP072312-01) were determined as 51.2% and 47.2% respectively, with atotal content of 98.4% in the extract.

The natural content of honokiol and magnolol in Magnolia bark werereported in a range of 2-11% and 0.3-4.6% respectively. The totalcontent of honokiol and magnolol in the commercial Magnolia bark extractcould be enriched and customized to a range of 1-99%.

Example 8 Anti-Microbial Activities of Magnolia Stem Bark Extracts

Magnolia bark with a common name houpu in Chinese is one of the populartraditional herbal medicines with a very wide range of applications.Many of the formulations with Magnolia bark are used in treating lungdiseases such as including cough and asthma or intestinal infections andspasms, relieving abdominal swelling of various causes and edema.Essential oil was identified as the active constituents with two majormarker compounds honokiol and magnolol. Modern researches have shownthat Magnolia stem bark extract exhibited potent anti-microbialactivities. Seven microorganisms were chosen to test the anti-microbialactivity and Magnolia's potential usage as anti-microbial agentsincluding bacterial Staphylococcus aureus, Escherichia coli, and fungusstrains Microsporum canis, Trichophyton mentagrophytes, Trichophytonrubrum, A. brasiliensis and C. albicans. MIC and MBC were determinedfollowing the standard protocol as described in example 6 with activityresults in Table 6. Except anti-bacterial activity, Magnolia stem barkExtract also demonstrated d potent anti-fungal activity with a MIC valueof 80 ug/mL against C. albicans.

TABLE 6 Anti-microbial activities of Magnolia stem bark ExtractFP072312-01 Organism/strain MIC (ug/mL) MBC (ug/mL) Staphylococcusaureus 80 * Escherichia coli negative negative Pseudomonas aeruginosanegative negative Candida albicans 80 * Aspergillus brasiliensis 400  *Microsporum canis * 118  Trichophyton mentagrophytes 59 59 Trichophytonrubrum 59 59 * Unable to be determined

Example 9 Anti-Fungal Activity of the Mixture of Albizia Seeds Extractand Magnolia Bark Extract

In order to find composition effective against all five microorganismsrequired by category 2 preserve in USP chapter <51>, mainly targeting onboosting the anti-fungal activity, specifically against C. albicans andA. brasiliensis, the Albizia amara seeds extract and Magnolia barksextract were blended together to search for unexpected enhancedoutcomes.

These two materials were formulated at different ratios by mixing thesamples dissolved in methanol with desired amount and then dried invacuum to get the mixture. By manufacture, the Albizia amara seedsextract and Magnolia barks extract could be directly mixed at thedesired ratio to prepare the combination composition.

Compositions 1A:1M and 1A:3M, where A stands for Albizia amara seedsextract and M for Magnolia barks extract, yielded similar inhibitionswith MIC at 30 ug/mL against C. albicans, relatively weaker activityagainst A. brasiliensis with MIC at 130 ug/m L. The third ingredientPsoralea corylifolia seeds extract (P) was added to the 1A: 6M compo ata ratio of 3P, however this three-ingredient formulation (1A:6M:3P)didn't show superior activity compared with two-ingredient compositions.Given the fact that the 1:1 ratio of the Albizia amara seeds extract andMagnolia barks extract composition (416-145-50) resulted in the best MICagainst both C. albicans and A. brasiliensis. This composition wasconsidered as the lead composition and was selected for furtherassessment in the anti-microbial effectiveness test following <USP51>chapter with details in Example 12.

TABLE 7 Anti-microbial activities of combinations against C. albicansand A. brasiliensis. Candida albicans Aspergillus brasiliensis Samplecode Composition MIC (ug/mL) MIC (ug/mL) 416-145-50 A:M = 1:1 30 130416-145-75 A:M = 1:3 30 130 416-145-M3 A:M:P = 1:6:3 60 130 * A: Albiziaamara seeds extract. M: Magnolia barks extract. P: Psoralea corylifoliaseeds extract

Example 10 Evaluations of Synergy for the Compositions of Mixture ofAlbizia Seed Extracts and Magnolia Bark Extract

In vitro antifungal combinations are commonly evaluated on the basis ofthe fractional inhibitory concentration (FIC) index, which representsthe sum of the FIC of each substance tested. FIC is defined for eachsubstance as (MIC of drug A in combination/MIC of drug A alone). The FICindex equation is based on the hypothesis that that a drug cannotinteract with itself and therefore the effect of a self-drug combinationwill be always additive with FIC index at 1. The FIC index lower orhigher than 1 indicates synergy or antagonism in terms of theinteractions between two drugs combined. Considering the 2-fold drugdilution scheme, FIC index in the range of 0.5-4 was considered nointeraction either additive or indifference for the drug combinations.Cutoff of 0.5 or less was suggested for defining synergy, while cutoff 4or above was interpreted as antagonism (Farrar 1973; Huang 2013; Odds,2003; Pankey, 2005).

The fractional inhibitory concentration (FIC) was calculated with theequation: FIC of drug A (FIC A)=MIC of drug A in combination/MIC of drugA alone, and FIC of drug B (FIC B)=MIC of drug B in combination/MIC ofdrug B alone]. The FIC Index (FICI), calculated as the sum of each FIC(ΣFIC). The FIC index for the combination study was calculated as shownin Table 8. The calculated index of 1A:1M and 1A:3M against C. albicanswere 0.187 and 0.292. For A. brasiliensis, similar calculation leads tothe FIC index at 0.293 and 0.309 for 1A:1M and 1A:3M respectively. TheFCI index at 0.187 and 0.282 for the 1A:1M combination showed greatercertainty as synergistic effect instead of additive. For 1A:3Mcombination, similar synergistic effect was observed against these twofungal strains with FIC index at 0.282 and 0.309. 1A:6M:3P combinationhad a weak synergistic effect against C. albican—but no interactioneffect against A. brasiliensis.

The anti-fungal activity of this combination study indicated theexistence of synergy in formulating these two ingredients at a specificratio. The combination of 1A:1M was found with boosted anti-fungaleffects against both C. albicans and A. brasiliensis, which wasclassified as synergistic interaction based on the FIC index analysiswith FIC index less than 0.5. The merit of putting the Albizia amaraseeds extract and Magnolia barks extract together was confirmed by theirunexpected enhanced anti-fungal activity against both C. albicans and A.brasiliensis.

TABLE 8 Unexpected synergistic anti-fungal activity of composition ofAlbizia amara seeds extract and Magnolia stem barks extract AspergillusCandida albicans brasiliensis MIC MIC Sample code Composition (ug/mL)FIC index (ug/mL) FIC index A negative 500 M 80 400 P negative 30416-145-50 A:M = 1:1 30 0.188 130 0.293 416-145-75 A:M = 1:3 30 0.282130 0.309 416-145-M3 A:M:P = 1:6:3 60 0.45 130 1.521 * A: Albizia amaraseeds extract. M: Magnolia barks extract. P: Psoralea corylifolia seedsextract

Example 11 Effectiveness Test of Mixture of Albizia Seed Extract andMagnolia Bark Extract

The antibacterial activity and efficacy of a contemplated preservativewas evaluated following the USP <51> guideline. The USP <51> test methodis a quantitative assessment of antimicrobial preservatives added tonon-sterile dosage forms. The parameters used for this test aredetermined by the nature product and which of 4 categories it appliesto. Typical contact times extend to 28 days with observations inremaining microbial content made in 7 day intervals. This test istraditionally accompanied by a neutralization validation assay toconfirm the neutralization method employed is appropriate for the testsubstance and microorganism. Test substance 1A:1M combination(416-145-50) was dissolved in DMSO at a concentration of 1 mg/mL. Theresults are shown in FIG. 3. The test microorganisms Staphylococcusaureus (ATCC 6538), Escherichia coli (ATCC 8739), Pseudomonas aeruginosa(ATCC 9027), Candida albicans (ATCC 10231), Aspergillus brasiliensis(ATCC 16404) are prepared by growth in liquid or on agar culture medium.Microorganisms grown in liquid culture are centrifuged and washed priorto test. Suspensions of test microorganisms are standardized by dilutionin a buffered saline solution. Test and control substances aredispensed, in similar known volumes, to sterile vessels. Independentvolumes of Test and Control substances are inoculated with each testmicroorganism mixed and incubated. Control substances are immediatelyharvested and represent the concentration present at the start of thetest, or time zero.

Incubated Test Substances are harvested at the conclusion of eachcontact time by chemical neutralization. The number of survivingmicroorganisms are assessed at 2, 7, 14, and 28 days and logarithmicreductions are calculated based on initial concentrations observed attime zero. The Log₁₀ Reduction was calculated as Log (B/A), whereB=Number of viable test microorganisms in the control substancesimmediately after inoculation, and A=Number of viable testmicroorganisms in the test substances after the contact time. 416-145-50significantly reduce the counts of P. aeruginosa, and A. brasiliensiswith Log₁₀ reduction at 5.58 and 4.1 respectively and significantly, ifnot completely, eliminate S. aureus, E. coli, and C. albicans after 2days, and no bacteria or fungi growth was observed after 7 days, 14days, and 28 days against all five microorganisms. This study clearlydemonstrated that the effectiveness of this composition 416-145-50against E. coli, P. aeruginosa, S. aureus, A. brasiliensis, and C.albicans and meet the category 2 requirement which are no less than2-Log₁₀ reduction from the initial count 14 days and no increase fromthe 14 day's count at day 28 for bacteria and no increase from theinitial calculated count at 14 and 28 days for yeast and molds.

TABLE 9 Colony counts and Log reduction for 416-145-50 at 1 mg/mL TestMicroorganism P. A. C. Contact Data E. coli aeruginosa S. aureusbrasiliensis albicans Time Description 8739 9027 6538 16404 10231 TimeCFU/mL  2.60E+05  3.80E+06  3.55E+05  1.27E+05  6.10E+05 Zero 2 DaysCFU/mL <5.00E+00  1.00E+01 <5.00E+00  1.00E+01 <5.00E+00 Log₁₀ >4.72 5.58 >4.85 4.1 >5.09 Reduction 7 Days CFU/mL <5.00E+00 <5.00E+00<5.00E+00 <5.00E+00 <5.00E+00 Log₁₀ >4.72 >5.88 >4.85 >4.40 >5.09Reduction 14 Days  CFU/mL <5.00E+00 <5.00E+00 <5.00E+00 <5.00E+00<5.00E+00 Log₁₀ >4.72 >5.88 >4.85 >4.40 >5.09 Reduction 28 Days  CFU/mL<5.00E+00 <5.00E+00 <5.00E+00 <5.00E+00 <5.00E+00Log₁₀ >4.72 >5.88 >4.85 >4.40 >5.09 Reduction Note: The limit ofdetection for this assay is 5.00E+00 CFU/mL. Samples with no microbialrecovery are reported as <5.00E+00 in the chart.

Example 12 Effectiveness Test of Mixture of Albizia Seed Extract andMagnolia Bark Extract

The antibacterial activity and efficacy of 1A:1M combination(416-145-50) was further evaluated following the USP <51> guideline asdescribed in example 12 at a concentration of 0.5 mg/mL against E. coli,P. aeruginosa, S. aureus, A. brasiliensis, and C. albicans. The resultsare shown in FIG. 4. The number of surviving microorganisms are assessedat 48 hours, 7, 14, and 28 days and logarithmic reductions arecalculated based on initial concentrations observed at time zero asshown in Table 10. This study clearly demonstrated that theeffectiveness of this composition 416-145-50 at 0.05% against all fivemicroorganism E. coli, P. aeruginosa, S. aureus, A. brasiliensis, and C.albicans and meet the category 2 requirement which are no less than2-Log₁₀ reduction from the initial count 14 days and no increase fromthe 14 day's count at day 28 for bacteria and no increase from theinitial calculated count at 14 and 28 days for yeast and molds.

TABLE 10 Colony counts and Log reduction for 416-145-50 at 500 ug/mLTest Microorganism P. A. C. Contact Data E. coli aeruginosa S. aureusbrasiliensis albicans Time Description 8739 9027 6538 16404 10231 TimeCFU/mL  4.30E+05  2.40E+05  2.45E+05 2.50E+05  3.20E+06 Zero 2 DaysCFU/mL <5.00E+00 <5.00E+00 <5.00E+00 1.00E+01 <5.00E+00Log₁₀ >4.93 >4.68 >4.69 4.4 >5.81 Reduction 7 Days CFU/mL <5.00E+00<5.00E+00 <5.00E+00 1.00E+01 <5.00E+00 Log₁₀ >4.93 >4.68 >4.69 4.4 >5.81Reduction 14 Days  CFU/mL <5.00E+00 <5.00E+00 <5.00E+00 <5.00E+00 <5.00E+00 Log₁₀ >4.93 >4.68 >4.69 >4.70 >5.81 Reduction 28 Days  CFU/mL<5.00E+00 <5.00E+00 <5.00E+00 <5.00E+00  <5.00E+00Log₁₀ >4.93 >4.68 >4.69 >4.70 >5.81 Reduction Note: The limit ofdetection for this assay is 5.00E+00 CFU/mL. Samples with no microbialrecovery are reported as <5.00E+00 in the chart.

Example 13 Antimicrobial Activity of Mixture of Albizia Seed Extract andMagnolia Bark Extract

Minimum Inhibitory Concentration against bacteria S. aureus, P.aeruginosa, and E. coli, were determined for 1A:1M combination ofAlbizia amara seeds extract and Magnolia bark extract. The MinimumInhibitory Concentration Method (MIC) is carried out following thegeneral microbiological protocol to measure the lowest level of anantimicrobial agent that can inhibit microbial proliferation in liquidas described in example 6 with activity results in Table 11. Potentantimicrobial activity against Staphylococcus aureus was found with MICof 20 ug/mL. MIC of 1000 ug/mL was determined against both gram negativebacteria Escherichia coli and Pseudomonas aeruginosa for this Albiziaamara seeds extract and Magnolia bark extract combination.

TABLE 11 Anti-microbial activities of 1A:1M combination (416-145-50)Organism/strain MIC (ug/mL) Staphylococcus aureus 20 Escherichia coli1000 Pseudomonas aeruginosa 1000

Example 14 Antimicrobial Activity of Albizia Amara Seeds Extract AgainstEscherichia Coli at PH=3

Minimum Inhibitory Concentration against gram negative bacteriumEscherichia coil at acidic condition (PH=3) was determined for Albiziaamara seeds extract (RN368-31-mix).

The Minimum Inhibitory Concentration Method (MIC) is carried outfollowing the general microbiological protocol to measure the lowestlevel of an antimicrobial agent that can inhibit microbial proliferationin liquid. The test substance is dissolved in buffer solution at PH=3with mixture of citric acid and sodium citrate in water, then preparedby several serial 1:1 dilutions in a 96-well microtiter plate, throughMueller-Hinton broth or other appropriate medium. All wells or tubescontaining diluted test substances are inoculated with testmicroorganisms, individually, resulting in one additional and finaldilution of the product in all test vessels. The microtiter plate isincubated for 18-24 hours. After the incubation period, observations aremade to determine the minimum concentration (MIC) which is the lowestconcentration of test substance in a well where no turbidity (indicativeof test microorganism growth) is observed. The MIC was 125 ug/mL againstE. coli for the Albizia amara seed extract at PH=3, slightly weakeractivity compared to MIC at neural condition (62.5 ug/mL).

Example 15 Antifungal Activity of Albizia Amara Seeds Extract andMagnolia Bark Extract Combination Against Aspergillus Brasiliensis atpH=3

Minimum Inhibitory Concentration against A. brasiliensis at acidiccondition (PH=3) was determined for the Albizia amara seeds extractMagnolia bark extract combination.

The Minimum Inhibitory Concentration Method (MIC) is carried outfollowing the general microbiological protocol to measure the lowestlevel of an antimicrobial agent that can inhibit microbial proliferationin liquid. The test substance is dissolved in buffer solution at PH=3with mixture of citric acid and sodium citrate in 20% DMSO/water, thenfurther prepared by several serial 1:1 dilutions in a 96-well microtiterplate, through Mueller-Hinton broth or other appropriate medium. Allwells or tubes containing diluted test substances are inoculated withtest microorganisms, individually, resulting in one additional and finaldilution of the product in all test vessels. The microtiter plate ortest tubes are incubated for 18-24 hours. After the incubation period,observations are made to determine the minimum concentration (MIC) whichis the lowest concentration of test substance in a well where noturbidity (indicative of test microorganism growth) is observed. The MICwas determined as 125 ug/mL for this 1A:1M combination at PH=3. Theantifungal activity is improved for this 1A:1M combination at acidiccondition compared to original MIC of 400 ug/mL.

Example 16 Formulation of Mixture of Albizia Seed Extract and MagnoliaBark Extract

1A:1M combination (B1-11202015) was further formulated with non-ionico/w emulsion system at two concentrations 0.1% and 0.5% respectively. 3%(% by weight) 1,2-hexanediol was used as carrier solvent for B1-11202015by weight in cream formulations with PH of the emulsion in a range of6.0-6.5. B1-11202015 was completely dispersed in 3% 1,2 Hexanediol byweight at 50° C. before adding to the cream base. The cream base wasprepared by mixing all ingredients at 70° C. until it is uniform and allsolid materials have melted and mixed. The PH value of the cream basematerial will be adjusted to 6.0-6.5 after cooling down to 60° C. byadding 20% NaOH solution or 20% citric acid solution. The cream base wascool down to 50° C. before adding the 3% 1,2 Hexanediol and B1-11202105.The PH value of the formulated material will be further adjusted to therange of 6.0-6.5 if necessary. Two formulated samples were prepared with0.5% 1A:1M combination (1940601) and 0.1% 1A:1M (1940602) in the samefashion.

Example 17 Effectiveness Test of Formulated Albizia Seed Extract andMagnolia Bark Extract Combination

The antibacterial activity and efficacy of non-ionic o/w emulsionformulated 1A:1M combination at a concentration of 0.5% (1940601) wasevaluated following the USP <51> guideline as described in Example 11against E. coli, P. aeruginosa, S. aureus, A. brasiliensis, and C.albicans. The results are shown in FIG. 5. The number of survivingmicroorganisms is assessed at 2, 7, 14, and 28 days and logarithmicreductions are calculated based on initial concentrations observed attime zero as shown in Table 12. This study clearly demonstrated theeffectiveness of this composition 1940601 at 0.5% against all threebacteria, E. coli, P. aeruginosa, S. aureus, and the yeast C. albicans,with no bacteria or fungi growth observed after 2 days, and extend to 7days, 14 days, and 28 days. For A. brasiliensis, 1940601 showedrelatively weaker inhibition with Log₁₀ reduction 0.73 and 1.53 at D2and D7 respectively. It did reach 2-LOG₁₀ reduction at D14 (2.32) at D28(3.32).

This study clearly demonstrated that the effectiveness of thiscomposition 1940601 against all five strains E. coli, P. aeruginosa, S.aureus, A. brasiliensis, and C. albicans as category 2 products bypassing USP51 criteria, which require no less than 2-LOG₁₀ reductionfrom the initial count at 14 days, and no increase from the 14 dayscount at 28 days for bacteria and no increase from the initial count at14 and 28 days for yeast and mold.

TABLE 12 Colony counts and Log reduction for 1940601 at 0.5%concentration Test Microorganism P. A. C. Contact Data E. coliaeruginosa S. aureus brasiliensis albicans Time Description 8739 90276538 16404 10231 Time CFU/mL  5.83E+05  1.90E+05  3.71E+05 1.04E+05 3.30E+05 Zero 2 Days CFU/mL <5.00E+01 <5.00E+01 <5.00E+01 1.95E+04<5.00E+01 Log₁₀ >4.03 >3.58 >3.87 0.73 >3.82 Reduction 7 Days CFU/mL<5.00E+01 <5.00E+01 <5.00E+01 3.05E+03 <5.00E+01 Log₁₀ >4.03 >3.58 >3.871.53 >3.82 Reduction 14 Days  CFU/mL <5.00E+01 <5.00E+01 <5.00E+015.00E+02 <5.00E+01 Log₁₀ >4.03 >3.58 >3.87 2.32 >3.82 Reduction 28 Days CFU/mL <5.00E+01 <5.00E+01 <5.00E+01 5.00E+01 <5.00E+01Log₁₀ >4.03 >3.58 >3.87 3.32 >3.82 Reduction Note: The limit ofdetection for this assay was 50 CFU/g. Values observed below the limitof detection are noted as <5.00E+01 in the table.

Example 18 Effectiveness Test of Formulated Albizia Seed Extract andMagnolia Bark Extract Combination

The antibacterial activity and efficacy of 1A:1M combination formulatedwith non-ionic o/w emulsion system (1940602) was further evaluated at0.1% concentration in the challenge testing following USP <51> guidelineas described in Example 11 against 5 strains E. coli, P. aeruginosa, S.aureus, A. brasiliensis, and C. albicans. The results are shown in FIG.6. The number of surviving microorganisms is assessed at 2, 7, 14, and28 days and logarithmic reductions are calculated based on initialconcentrations observed at time zero as shown in Table 13.

This study demonstrated that this composition 1940602 at 0.1% eliminatedE. coli, P. aeruginosa, S. aureus, and C. albicans starting Day 2. Nogrowth was observed at all four contact time points D2, D7, D14 and D28.For A. brasiliensis, no inhibition was found at D2, with improvementafter D2. Log₁₀ reductions are 1.02, 0.23 and 1.33 at D7, D14, and D28respectively. This study clearly showed that this composition 1940602was effective against E. coli, P. aeruginosa, S. aureus, A.brasiliensis, and C. albicans and qualified 1940602 as category 2products by passing USP51 criteria, which require no less than 2-LOG₁₀reduction from the initial count at 14 days, and no increase from the 14day's count at 28 days for bacteria and no increase from the initialcount at 14 and 28 days for yeast and mold.

TABLE 13 Colony counts and Log reduction for 1940602 at 0.1%concentration Test Microorganism P. A. C. Contact Data E. coliaeruginosa S. aureus brasiliensis albicans Time Description 8739 90276538 16404 10231 Time CFU/mL  8.55E+05  6.70E+05  6.45E+05 4.35E+05 3.40E+05 Zero 2 Days CFU/mL <5.00E+01 <5.00E+01  5.00E+01 7.00E+05<5.00E+01 Log₁₀ >4.23 >4.13  4.11 None >3.83 Reduction 7 Days CFU/mL<5.00E+01 <5.00E+01 <5.00E+01 4.20E+04 <5.00E+01 Log₁₀ >4.23 >4.13 >4.111.02 >3.83 Reduction 14 Days  CFU/mL <5.00E+01 <5.00E+01 <5.00E+012.55E+05 <5.00E+01 Log₁₀ >4.23 >4.13 >4.11 0.23 >3.83 Reduction 28 Days CFU/mL <5.00E+01 <5.00E+01 <5.00E+01 2.55E+04 <5.00E+01Log₁₀ >4.23 >4.13 >4.11 1.33 >3.83 Reduction Note: The limit ofdetection for this assay was 50 CFU/g. Values observed below the limitof detection are noted as <5.00E+01 in the table.

REFERENCES

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Thus, specific embodiments and methods of compounds and compositionsuseful for preservatives and antimicrobial agents, includingstereoisomers, pharmaceutically or nutraceutically acceptable salts,tautomers, glycosides and prodrugs of the disclosed compounds, alongwith related methods of improving and maintaining liver health have beendisclosed. It should be apparent, however, to those skilled in the artthat many more modifications besides those already described arepossible without departing from the inventive concepts herein. Theinventive subject matter, therefore, is not to be restricted except inthe spirit of the disclosure herein. Moreover, in interpreting thespecification and claims, all terms should be interpreted in thebroadest possible manner consistent with the context. In particular, theterms “comprises” and “comprising” should be interpreted as referring toelements, components, or steps in a non-exclusive manner, indicatingthat the referenced elements, components, or steps may be present, orutilized, or combined with other elements, components, or steps that arenot expressly referenced.

The invention claimed is:
 1. An antimicrobial composition, comprising: amixture of at least one Albizia extract and a Magnolia extract, whereinthe at least one Albizia extract is enriched for one or more macrocyclicalkaloids and wherein the at least one Magnolia extract is enriched forone or more lignans, wherein the combination of the at least one Albiziaextract and a Magnolia extract is present in a range from about 2.5% toabout 10% by weight of the total weight of the composition, and whereinthe Albizia extract and the Magnolia extract are blended in a weightratio ranging from 3:1 to 1:3.
 2. A food, cosmetic, nutraceutical orpharmaceutical that comprises the composition of claim 1, wherein thecomposition is a preservative.
 3. The composition of claim 2, whereinthe composition is present in the food, cosmetic, nutraceutical orpharmaceutical in a range from about 0.001% to about 10% by weight ofthe total weight of the food, cosmetic, nutraceutical or pharmaceutical.4. The composition of claim 3, wherein the composition is present in thefood, cosmetic, nutraceutical or pharmaceutical in a range from about0.01% to about 5% by weight of the total weight of the food, cosmetic,nutraceutical or pharmaceutical.
 5. The composition of claim 1, whereinthe Albizia extract and the Magnolia extract are blended in a weightratio at 1:1.
 6. The composition of claim 1, wherein the Albizia extractcomprises about 0.01% to 99.9% of macrocyclic alkaloids.
 7. Thecomposition of claim 1, wherein Magnolia extract comprises about 0.01%to 99.9% of diphenol lignans.
 8. The composition of claim 1, wherein theAlbizia comprises Albizia amara.
 9. The composition of claim 1, whereinthe at least one enriched Albizia extract is extracted from Albiziaplants with water, ethanol, methanol, alcohol and water-mixed solvents,or by supercritical fluid.
 10. The composition of claim 1, wherein thealkaloids isolated from the at least one Albizia extract compriseBudmunchiamine A, B, C, D, E, F, G, H, I, J, K, Budmunchiamine L1, L2,L3, L4, L5, L6, 13-Normethylbudmunchiamine K, 9-Normethylbudmunchiam ineK, 1-Normethylbudmunchiamine K, 6′-Hydroxy-9-normethylbudmunchiamine K,6′-Hydroxybudmunchiamine K or a combination thereof.
 11. The compositionof claim 1, wherein the at least one enriched Magnolia extract isextracted from Magnolia plants with water, ethanol, methanol, alcoholand water mixed solvents, or by supercritical fluid.
 12. The compositionof claim 1, wherein the Magnolia comprises Magnolia officinalis.
 13. Thecomposition of claim 1, wherein the lignans isolated from the at leastone Magnolia extract comprise magnolol, honokiol or a combinationthereof.
 14. The composition of claim 1, wherein the compositioncomprises: a mixture of at least one Albizia extract from a plantmaterial or other source, and at least one Magnolia extract from a plantmaterial or another source, wherein the at least one Albizia extract hasan increase in an amount or an activity of one or more macrocyclicalkaloids as compared to the amount or the activity of the one or moremacrocyclic alkaloids in the plant material or other source of the atleast one Albizia extract, and wherein the at least one Magnolia extracthas an increase in an amount or an activity of one or more lignans ascompared to the amount or the activity of the one or more lignans in theplant material or other source of the at least one Magnolia extract. 15.The composition of claim 14, wherein the plant material or other sourcecomprises stems, stem barks, trunks, trunk barks, twigs, tubers, roots,root barks, young shoots, seeds, rhizomes, flowers and otherreproductive organs, tissue cultures and stem cell cultures, leaves andother aerial parts or a combination thereof.
 16. The composition ofclaim 1, wherein the composition further comprises a pharmaceutically ornutraceutically acceptable carrier, diluent, or excipient, wherein thepharmaceutical or nutraceutical formulation comprises from about 0.001weight percent (wt %) to about 10 wt % of active ingredients of theextract mixture.
 17. The composition of claim 1, wherein the compositionis formulated as a tablet, hard capsule, or soft gel capsule.
 18. Thecomposition of claim 1, wherein the composition additionally comprisesat least one known antimicrobial ingredient.
 19. The composition ofclaim 18, wherein the composition additionally comprises at least oneknown antimicrobial ingredients, wherein the at least one knownantimicrobial ingredient comprises plant powder or plant extract ofPsoralea corylifolia.
 20. The composition of claim 18, wherein thecomposition additionally comprises at least one known antimicrobialingredients, wherein the at least one known antimicrobial ingredientcomprises calcium propionate.